Capsaicin compound and preparation method and application thereof

Abstract

The invention discloses a capsaicin compound and a preparation method and application thereof, the molecular formula of capsaicin Capsivaryine is C16H21NO3, the chemical structure of capsaicin Capsivaryine is as follows, and the capsaicin Capsivaryine compound is a new compound reported for the first time and is prepared by taking mature and dry fruits of capsicum as raw materials through crushing, sieving, silica gel column treatment, D101 macroporous resin treatment and HPLC (High Performance Liquid Chromatography). The compound disclosed by the invention has a certain in-vitro inhibition effect on human breast cancer cells MCF-7, human liver cancer cells HEPG2, human non-small cell lung cancer cells A549 and human ovarian cancer cells SKOV3, and has no toxicity on human normal liver cells L02. The compound is a natural product which is novel in structure and good in activity, has great potential in the anti-tumor aspect, enriches diversity of pepper plants, and provides guidance for research of chemical components of pepper. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a capsivaryine compound and a preparation method and application thereof. Background Art

[0002] Pepper (Capsicum annuum) is an annual or limited perennial herb of the genus Capsicum in the family Solanaceae. It is widely used as a medicinal and edible plant in the world. The spicy ingredients in pepper fruits are mainly composed of a series of substances that are very similar in structure and properties, which are collectively called capsaicinoids.

[0003] Capsaicin has high biological activity, such as anti-tumor, anti-inflammatory, antioxidant, bactericidal, insecticidal, etc. It is widely used in medicine, food, military industry and other fields, and has high social and economic benefits, and has always been a hot topic of research at home and abroad.

[0004] However, there are only more than 20 natural capsaicinoids discovered so far, of which capsaicin and dihydrocapsaicin account for about 90% of the capsaicinoid content, and the content of five types of capsaicinoids is close to 99%. It is difficult to separate other types of capsaicinoids. Natural products are the main source of oral drugs. Some existing drugs cause cumulative toxicity and high prices after long-term use. New drug research and development is needed to address the limitations of existing drugs. Summary of the invention

[0005] Purpose of the invention: In view of the problems existing in the prior art, the present invention provides a novel capsivaryine compound, which has high activity against various tumor cells and is non-toxic to normal cells, and can be used to prepare anti-tumor drugs.

[0006] The invention also provides a preparation method and application of the compound.

[0007] Technical solution: To achieve the above purpose, the present invention provides a capsaicin Capsivaryine, wherein the molecular formula of the capsivarycin Capsivaryine is C 16 H 21 NO3, its chemical structure is as follows:

[0008]

[0009] Wherein, the capsaicin is separated from Capsicum annuum.

[0010] The preparation method of the capsaicin compound of the present invention comprises the following steps:

[0011] (1) crushing the dried chili pepper fruits, then soaking them in an organic solvent, concentrating the filtrates under reduced pressure and combining them to obtain a chili pepper extract;

[0012] (2) immersing the extract in water, and extracting with petroleum ether, ethyl acetate, and n-butanol in sequence to obtain a petroleum ether extract, an ethyl acetate extract, and an n-butanol extract;

[0013] (3) The ethyl acetate extract was separated into Fr 1, Fr 2, Fr 3, Fr 4, Fr 5 and Fr 6 components by stepwise elution with a silica gel column in a petroleum ether-ethyl acetate gradient system;

[0014] (4) Fr5: This component was further separated by stepwise elution using a silica gel column in a dichloromethanol-methanol gradient system to obtain seven components, Fr5.1, Fr5.2, Fr5.3, Fr5.4, Fr5.5, Fr5.6, and Fr5.7;

[0015] (5) Fr5.2 was depigmented and then gradient eluted on silica gel with petroleum ether-ethyl acetate to obtain Fr5.2.1, Fr5.2.1, Fr5.2.2, Fr5.2.3, Fr5.2.4, Fr5.2.5, Fr5.2.6, Fr5.2.7, Fr5.2.8, Fr5.2.9, Fr5.2.10, and Fr5.211;

[0016] (6) Fr5.2.10 was analyzed by semi-preparative HPLC and purified to obtain the target compound Capsivaryine.

[0017] Wherein, the organic solvent in step (1) is ethanol, and its volume concentration is 80-85%; the soaking time is 7-10 days, and the soaking temperature is 40-50° C.; the soaking and reduced pressure concentration of the filtrate are repeated 4-6 times.

[0018] Wherein, in step (2), petroleum ether, ethyl acetate and n-butanol are used for extraction in sequence, and each extraction is repeated 3-5 times.

[0019] Wherein, in step (3), the petroleum ether-ethyl acetate gradient system has a separation volume ratio of 15:1-1:3 and is eluted in sequence, that is, the solvent ratio is gradually reduced from 15:1, 14:1, 13:1, etc.

[0020] Wherein, the separation condition in step (4) is a dichloromethanol-methanol gradient system, and the separation volume ratio is 50:1, 40:1, 30:1, 20:1, 10:1, 9:1-1:1 in sequence for gradient elution, that is, the solvent ratio gradually decreases from 50:1, 40:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1, etc.

[0021] In step (5), the pigment is removed by using ethanol on D101 macroporous resin, the separation conditions are petroleum ether-ethyl acetate system, the separation volume ratio is 7:1-1:1 in sequence, that is, the solvent ratio is gradually reduced from 7:1, 6:1, 5:1, etc.

[0022] Preferably, the silica gel column used in steps (3), (4), and (5) is 200-300 mesh.

[0023] Wherein, the separation conditions of the semi-preparative high performance liquid chromatography in step (6) are acetonitrile-water as the mobile phase, isocratic elution is performed at a volume ratio of 65:35 to 70:30, the flow rate is 2-3 mL / min, the temperature is controlled at 35-45° C., and each elution time is 30-45 minutes. The target compound elutes within 8-12 minutes, and is collected in sections according to the elute time and combined and concentrated.

[0024] The invention relates to the use of the capsaicin compound in the preparation of anti-tumor drugs.

[0025] Wherein, the tumor is breast cancer, liver cancer, non-small cell lung cancer, and ovarian cancer.

[0026] Preferably, the tumor cells are human breast cancer cells MCF-7, human liver cancer cells HEPG2, human non-small cell lung cancer cells A549, and human ovarian cancer cells SKOV3.

[0027] The pharmaceutical composition for preventing or treating tumors of the present invention contains capsivaryine or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0028] The pharmaceutical composition of the present invention is used in the preparation of anti-tumor drugs, wherein the tumors are human breast cancer, liver cancer, non-small cell lung cancer, and ovarian cancer.

[0029] The compound described in the present invention is an alkaloid extracted from nature for the first time, and the compound is characterized by methods such as nuclear magnetic resonance, mass spectrometry, ultraviolet and infrared chromatography. The present invention uses mature dried fruits of pepper as raw materials, and prepares the capsivaryine compound through crushing and screening, silica gel column, D101 macroporous resin, HPLC structure and other routes, and determines the structure through modern spectral analysis. The obtained capsivaryine compound has high purity.

[0030] The present invention provides an alkaloid with a novel structure and a separation method, which enriches the diversity of the plant. The present invention separates the new capsaicinoids by traditional separation methods (such as HPLC, solid phase extraction, etc.), which may make the separation process more efficient and accurate, and improve the separation purity. Natural products are the main source of oral drugs, and the novel structure of the present invention may be used in the research and development of new drugs to solve the limitations of existing drugs.

[0031] The new compound in the present invention is based on the core principle of separation and purification of natural products, and the new compound can be extracted efficiently. The present invention separates the new compound from the capsaicinoid compound, and combines multiple separation techniques (chromatography and HCPT) to further improve the separation accuracy. The compound of the present invention is compared with the known capsaicin and HCPT, and is a compound of a completely new structure. The new compound has a significant effect of inhibiting tumor cells, and the inhibitory effect on MCF-7 cells is better than the above compounds, and the inhibitory effect on cells A549, HEPG2, and SKOV3 is significantly better than the compound N-vanillyl-4E,6E-dien-8-methylnonanamide.

[0032] The present invention is the first to find a double bond connected to the carbonyl group at the 1' position in the capsaicinoid compounds. Due to the novelty of the structure, the new capsivaryine compound has a stronger inhibitory effect on MCF-7 cells, especially stronger than the similar structural compound N-vanillyl-4E,6E-dien-8-methylnonanamide. The present invention expands the types of capsaicinoids by isolating new compounds, and provides a new reference for studying the relationship between the structure and function of capsaicinoids.

[0033] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0034] 1. The present invention proposes a novel capsivaryine compound, which has excellent in vitro inhibitory effects on human breast cancer cells MCF-7, human liver cancer cells HEPG2, human non-small cell lung cancer cells A549, and human ovarian cancer cells Skov3, and is non-toxic to normal human liver cells L02. JC-1 and apoptosis experiments show that the compound can induce apoptosis of MCF-7 cells according to the dose, and the compound may become a potential drug for the treatment of cancer diseases such as breast cancer.

[0035] 2. The preparation method of the capsivaryine compound of the present invention is simple and low in cost. Materials and equipment such as silica gel column chromatography and high performance liquid chromatography are used, and all reagents used can be recycled and reused. At the same time, silica gel separation is avoided, and the use of organic solvents such as chloroform and benzene is eliminated, which greatly reduces the pollution to the environment; at the same time, the clear structure also ensures that it can be obtained by artificial synthesis in the later stage. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Capsivaryine compounds 1 H NMR spectrum;

[0037] Figure 2 Capsivaryine compounds13 C NMR spectrum;

[0038] Figure 3 It is the spectrum of Capsivaryine compound DEPT135;

[0039] Figure 4 This is the high-resolution mass spectrum of Capsivaryine compound;

[0040] Figure 5 Capsivaryine compound HMBC and 1 H- 1 HCOSY correlation diagram;

[0041] Figure 6 The inhibitory capacity of CCK8 on four tumor cells and IC of MC7-7 cells 50 picture;

[0042] Figure 7 Schematic diagram of the JC-1 experimental results of mitochondrial membrane potential;

[0043] Figure 8 Schematic diagram of apoptosis experimental results. DETAILED DESCRIPTION

[0044] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0045] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources. Experimental methods without specific conditions specified in the examples are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0046] Example 1

[0047] Preparation of Capsivaryine Compound

[0048] (1) 20 kg of dried chili pepper fruits were crushed and then soaked in 80% ethanol solution at 40° C. for 7 days. The filtrate was collected and concentrated under reduced pressure. This process was repeated 4 times to obtain 2.6 kg of extract.

[0049] (2) The extract was immersed in 2.5 L of water and extracted with petroleum ether (10 L × 3 times), ethyl acetate (10 L × 3 times), and n-butanol (10 L × 3 times) in sequence. The organic phase was concentrated under reduced pressure to obtain 522 g of petroleum ether extract, 120 g of ethyl acetate extract, and 340 g of n-butanol extract.

[0050] (3) 120 g of ethyl acetate extract was eluted step by step with a 200-300 mesh silica gel column in a petroleum ether-ethyl acetate gradient system (15:1 to 1:3, v / v), and TLC spot plate merging was performed. The spot plate system was: petroleum ether-ethyl acetate (1:0 to 1:1, v / v), and Fr1 to Fr6 were obtained according to the polarity separation of the compounds (consistent color properties and close Rf values). Among them, Fr5 was separated by the above petroleum ether-ethyl acetate system (10:1-8:1, v / v). In the petroleum ether:ethyl acetate system (10:1, 9:1, 8:1, v / v), the separated samples were spot plated consistently with the petroleum ether-ethyl acetate system (5:1, v / v), and concentrated under reduced pressure at 40°C to obtain Fr5.

[0051] (4) Fr5 (39.1 g) This component was further chromatographed on a 200-300 mesh silica gel column using CH2Cl2 / methanol (50:1-1:1, v / v) and then TLC plated and combined. The plated system was petroleum ether-ethyl acetate (10:1 to 1:1, v / v). Seven components, Fr5.1-5.7, were separated according to the polarity of the compounds (consistent color properties and close Rf values). Among them, Fr5.2 was separated using the above CH2Cl2-methanol system (8:1-6:1, v / v). In the CH2Cl2:methanol system (8:1, 7:1, 6:1, v / v), the separated samples were plated consistently using the petroleum ether-ethyl acetate system (4:1, v / v), and concentrated under reduced pressure at 40°C to combine into Fr5.2.

[0052] (5) Fr5.2 (28 g) was depigmented with 80% ethanol on D101 macroporous resin, and the filtrate was concentrated under reduced pressure at 40°C, and then gradient eluted with petroleum ether / ethyl acetate (7:1 / 1:1, v / v) on a 200-300 mesh silica gel column to obtain Fr5.2.1-Fr5.2.11. Among them, Fr5.2.10 was separated by the above petroleum ether-ethyl acetate system (3:1-2:1, v / v). In the petroleum ether:ethyl acetate system (3:1, 2:1, v / v), the separated samples were plated with the petroleum ether-ethyl acetate system (2:1, v / v) to be consistent, and concentrated under reduced pressure and vacuum at 40°C to combine into Fr5.2.10.

[0053] (6) Fr5.2.10 (3 g) was analyzed by semi-preparative HPLC on a C18 column (250 mm × 10 mm, 5 μm) with acetonitrile-water as the mobile phase, and eluted isocratically at a volume ratio of 67:33, 3 mL / min, and 40°C. Each elution time was 40 min. The product was then distilled under reduced pressure and concentrated according to the peak elution time. The peak appeared in about 10 min. The compound capsivaryine was detected by full wavelength detection with a UV detector.

[0054] Example 2

[0055] Structural analysis of compounds

[0056] The compound capsivaryine of Example 1 is a brown oily solid. The molecular formula of the compound was determined by HRESIMS at m / z 276.1588 [M+H + ] is determined as C 16 H 21 NO3(C 16 H 21 The calculated value of NO3 is 276.1595), and the degree of unsaturation is 7. Capsivaryine compound 1 H NMR spectrum, 13 C NMR spectrum, DEPT135 spectrum and high-resolution mass spectrum are shown in Figure 1-4 shown.

[0057] Compound 13 C NMR spectroscopy ( Figure 2 ) showed 16 carbon signals, which were assigned to amide [δ C 166.2(C-1')], six aromatic carbons [δ C 130.4 (C-1) 110.9 (C-2), 146.8 (C-3) 145.3 (C-4) 114.5 (C-5) and 121.0 (C-6)], four olefinic carbons [δ C 121.7 (C-2'), 142.1 (C-3'), 125.4 (C-4') and 150.3 (C-5')], one methoxy carbon (dc 56.1), two methyl carbons δ C 22.0 (C-7', 8')], one methylene carbon [δ C 43.8(C-7)], and a methine carbon [δ C 31.6 (C-6')]. In the 1H NMR spectrum of 1, three aromatic protons [δ H 6.86 (1H, d = 8.0 Hz, H-5), 6.82 (1H, d, J = 1.9 Hz, H-2), 6.78 (1H, dd, J = 8.1, 1.9 Hz, H-6), so it is speculated that the compound contains a trisubstituted benzene ring and an amide functional group. At the same time, the above spectral data of the compound are compared with the known compound N-vanillyl-4E, 6E-dien-8-methylnonanamide, so it can be speculated that the compound is a capsaicinoid compound.

[0058] However, the double bond of the compound of the present invention is directly connected to the carbonyl group at the 1' position.

[0059]

[0060] At the same time, it was also observed that the two methyl groups [δ H 1.03 (d, J = 6.7 Hz, 6H, H-7', 8')], a methoxy group [δ H 3.88 (3H, s)], 1 methine proton [δ H 2.36~2.30(1H,m,H-6')] and one methylene proton [δ H 4.44 (2H, d, J = 5.7 Hz, H-7)] and [δ H 5.61 (2H, s, -NH, -OH)], double bonds at 2' and 3' [δ H 5.76(1H,d,J=15.0Hz,H-2')δ H 7.23 (1H, dd, J = 15.0, 9.7 Hz, H-3')], the coupling constant > 12 is judged to be a trans structure. The double bonds at the 4' and 5' positions [δ H 6.10-6.05 (2H, t, J = 8.8 Hz, H-4', H-5'), coupling constant < 12, judged to be a cis structure.

[0061] exist Figure 5 HMBC and 1 H- 1 In the HCOSY correlation diagram, it was observed that there were correlations between H-2' and C-1', C-4', H-5' and C-3'; H-8' to C-5'; H-7 to C-2, C-6; H-1 to C-5; H-6 to C-4 and H-OMe to C-3. The structure of the compound was thus determined and it was a new compound named Capsivaryine. The hydrogen and carbon spectrum data are shown in Table 1.

[0062] Table 1H (600MHz, CDCl3) and 13 C NMR (151 MHz, CDCl3) data (δ in ppm).

[0063]

[0064] 1) overlapping

[0065] Example 3

[0066] Cytotoxicity assay

[0067] The cells used in the experiment are internationally used tumor cells: human non-small cell lung cancer cells (A549), human liver cancer cells (HEPG2), human breast cancer cells (MCF-7), human ovarian cancer cells (SKOV3) and human normal liver cells (L02). The positive control drug is hydroxycamptothecin (HCPT), and the conventional CCK-8 method is used for testing.

[0068] Cell passaging: Once the cell density reaches 80%, wash the culture medium in the culture dish twice with PBS. Subsequently, add 1 mL of trypsin containing EDTA to digest the cells, followed by 4 mL of culture medium to terminate digestion. After repeated pipetting to form a single cell distribution, add the suspension together with its growth medium to a new culture dish and place it in a 37°C, 5% CO2 incubator for culture and expansion.

[0069] IC50 50 Determination

[0070] (1) Cells in the logarithmic growth phase were digested with trypsin, and culture medium was added to form a single cell suspension. The cells were counted using a counting plate. 5000 cells / well were inoculated in a 96-well plate for both the control group and the experimental group. 100 μL of culture medium was added to each well, and the 96-well plate was incubated in a 37°C, 5% CO2 incubator for 24 hours.

[0071] (2) Add a DMSO solution containing the capsivaryine compound, initially prepared at 1 mg / ml, and diluted with culture medium to change the concentration, so that the final DMSO content is less than 0.2%. Wash away the original culture medium in each well of the 96-well plate, and add 100 μL of culture medium containing different concentrations of the compound (3.125, 6.25, 12.5, 25, 50 μM) to each 96-well plate. Set up 3 replicates for each concentration, and incubate the 96-well plate in a 37°C, 5% CO2 incubator for 24 hours.

[0072] (3) Wash away the original culture medium in each well of the 96-well plate, add 100 μL of culture medium containing 10% CCK-8 solution to each well, the blank group is 100 μL of culture medium containing 10% CCK-8 solution without cells, and the control group is 100 μL of culture medium containing 10% CCK-8 solution with cancer cells, and incubate in an incubator for 1 hour. The OD value of a single well measured at 450 nm was measured by an ELISA instrument. The experiment was repeated three times for each compound. Calculated using GraphPad Prism 8.30 software. The half-maximal inhibitory concentration IC of the sample on tumor cells 50 The calculation results are shown in Table 2. Figure 6 As shown, the compound has a certain inhibitory effect on four tumor cells, and its inhibitory effect on MCF-7 cells is better than that of hydroxycamptothecin.

[0073] Table 2 Cytotoxicity of compounds, IC 50 (μM)

[0074]

[0075] The experimental results are shown in Table 2. The compounds of the present invention have certain in vitro inhibitory effects on human breast cancer cells MCF-7, human liver cancer cells HEPG2, human non-small cell lung cancer cells A549, and human ovarian cancer cells SKOV3, and have no toxicity to normal human liver cells L02. The inhibitory effect on MCF-7 breast cancer cells is better than HCPT, and the others are equivalent to HCPT.

[0076] The structure of HCPT is:

[0077] Example 4

[0078] Mitochondrial membrane potential JC-1 experiment

[0079] (1) Approximately 1×10 5 MCF-7 cells were treated with (0, 10, 15 and 20 μM) of the compound Capsivaryine for 4 hours in a confocal dish. 1 ml of the pre-prepared JC-1 staining solution and 1 ml of culture medium were added for staining, incubated in an incubator for 20 minutes, washed twice with the pre-prepared JC-1 buffer, the supernatant was aspirated, 2 ml of culture medium was added, and the intracellular mitochondrial membrane potential (MMP) level was detected by laser confocal microscopy.

[0080] (2) After the cells were treated with compounds at concentrations of 10, 15, and 20 μM for 4 hours, compared with the blank control group, the green fluorescence gradually increased, the red fluorescence gradually decreased, and the ratio of red to green fluorescence intensities decreased.

[0081] The downregulation of mitochondrial membrane potential (MMP) leads to early apoptosis, and the decrease in fluorescence ratio indicates that apoptosis is becoming more serious. Figure 7 As the drug concentration increases (10, 15, 20 μM), the red fluorescence gradually weakens and the red / green fluorescence intensity ratio decreases, reflecting an increase in early cell apoptosis. This indicates that capsaicin can cause a decrease in the mitochondrial membrane potential of MCF-7 cells, leading to cell apoptosis.

[0082] Example 5

[0083] Flow cytometry apoptosis assay

[0084] (1) Approximately 2.5×10 6MCF-7 cells were treated with different concentrations of the compound Capsivaryine (0, 10, 15 and 20 μM) in a six-well plate for 24 hours. The supernatant was added to a centrifuge tube, washed once with PBS, added to the centrifuge tube, digested with EDTA-free trypsin for 1 minute, digested with culture medium, added to the previous centrifuge tube, and centrifuged at 1000r for 5 minutes. Annexin V-FITC binding solution, Annexin V-FITC and propidium iodide staining solution were added together, incubated for 20 minutes, and detected by flow cytometry.

[0085] (2) Results Figure 8 ) showed that compared with the blank control group, the apoptosis rate was 17.07% at 10 μM and 32.08% at 20 μM. Therefore, capsaicin has an effect on inducing programmed cell apoptosis of MCF-7 cells in a dose-dependent manner.

[0086] In summary, the initial screening of cytotoxic activity, JC-1 experiment and apoptosis experiment showed that capsivaryine compounds inhibited the growth of MCF-7 and other cancer cells and induced their apoptosis. Capsaicin may become a potential drug for the treatment of breast cancer.

Claims

1. A capsaicin compound, characterized in that The capsaicin compound has the molecular formula C 16 H 21 NO3, its chemical structure is as follows:

2. The capsaicin compound according to claim 1, characterized in that The capsaicin is separated from Capsicum annuum.

3. A method for preparing the capsaicin compound according to claim 1, characterized in that: The steps include: (1) crushing the dried chili pepper fruits, then soaking them in an organic solvent, concentrating the filtrates under reduced pressure and combining them to obtain a chili pepper extract; (2) immersing the extract in water, and extracting with petroleum ether, ethyl acetate, and n-butanol in sequence to obtain a petroleum ether extract, an ethyl acetate extract, and an n-butanol extract; (3) The ethyl acetate extract was separated into Fr 1, Fr 2, Fr 3, Fr 4, Fr 5 and Fr 6 components by stepwise elution with a silica gel column in a petroleum ether-ethyl acetate gradient system; (4) Fr5: This component was further separated by stepwise elution using a silica gel column in a dichloromethanol-methanol gradient system to obtain seven components, Fr5.1, Fr5.2, Fr5.3, Fr5.4, Fr5.5, Fr5.6, and Fr5.7; (5) Fr5.2 was depigmented and then gradient eluted on silica gel with petroleum ether-ethyl acetate to obtain Fr5.2.1, Fr5.2.1, Fr5.2.2, Fr5.2.3, Fr5.2.4, Fr5.2.5, Fr5.2.6, Fr5.2.7, Fr5.2.8, Fr5.2.9, Fr5.2.10, and Fr5.211; (6) Fr5.2.10 was analyzed by semi-preparative HPLC and purified to obtain the target compound Capsivaryine.

4. The preparation method according to claim 3, characterized in that: The organic solvent in step (1) is ethanol, and its volume concentration is 80-85%; the soaking time is 7-10 days, and the soaking temperature is 40-50° C.; the soaking and reduced pressure concentration of the filtrate are repeated 4-6 times.

5. The preparation method according to claim 3, characterized in that: In step (2), petroleum ether, ethyl acetate and n-butanol are used for extraction in sequence, and each extraction is repeated 3-5 times.

6. The preparation method according to claim 3, characterized in that: In step (3), a petroleum ether-ethyl acetate gradient system is used, and the separation volume ratio is 15:1-1:3, and gradient elution is performed in sequence; in step (4), the separation conditions are a dichloromethanol-methanol gradient system, and the separation volume ratios are 50:1, 40:1, 30:1, 20:1, 10:1, 9:1-1:1, and gradient elution is performed in sequence; in step (5), the pigment is removed by using ethanol on a D101 macroporous resin, and the separation conditions are a petroleum ether-ethyl acetate system, and the separation volume ratio is 7:1-1:1, and gradient elution is performed in sequence.

7. The preparation method according to claim 3, characterized in that: The separation conditions of the semi-preparative HPLC in step (6) are as follows: the mobile phase is acetonitrile-water, the volume ratio is 65:35-70:30, the flow rate is 2-3 mL / min, the temperature is controlled at 35-45°C, each elution time is 30-45 min, the target compound elutes within 8-12 min, and the samples are collected in sections according to the elute time and combined and concentrated.

8. A pharmaceutical composition comprising the capsaicin compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 as an active ingredient and a pharmaceutically acceptable carrier.

9. Use of the capsaicin compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The tumor is preferably breast cancer, liver cancer, non-small cell lung cancer or ovarian cancer.

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